| Objective To explore the method of fixing GL13 K antimicrobial peptide on the surface of titanium dioxide(titanium dioxide)nanotubes and compare its effect,and select one of the best fixing methods to provide new ideas for the research of antimicrobial modification of titanium implant surface and the treatment of peri-implant inflammation.Methods A highly ordered porous structure of titanium nanotubes was constructed on the surface of pure titanium by anodic oxidation.The GL13 K antimicrobial peptide was immobilized on the surface of titanium nanotubes.The effects of several methods on the immobilization of antimicrobial peptides were compared by observing the surface morphology,composition and release kinetics of antimicrobial components.The antimicrobial coating on the surface of nanotubes was evaluated by in vitro antimicrobial experiments.Antibacterial effect of layer on Porphyromonas gingivalis.The specific experimental contents are as follows: 1.Treatment of Titanium Sheets and Preparation of Titanium Dioxide Nanotubes Round smooth titanium sheets with diameter of 15 mm were prepared.Porous structure of titanium nanotubes was constructed on the surface of titanium sheets by anodic oxidation.The morphology of titanium substrate was observed by FESEM.2.Surface loading of GL13 K antimicrobial peptide on titanium dioxide nanotubes Titanium nanotubes were used as blank control group(group A).GL13 K antimicrobial peptide was immobilized on the surface of titanium nanotubes by physical adsorption(group B),covalent binding(group C)and layer-by-layer self-assembly(group D).The four groups of samples were determined by contact angle measurement,Fourier transform infrared spectroscopy,X-ray photoelectron spectroscopy,positive fluorescence microscopy and ultraviolet spectrophotometer.The effect of GL13 K antimicrobial peptide immobilization on the surface of titanium nanotubes was analyzed.3.Inhibition of Porphyromonas gingivalis by titanium dioxide nanotubes loaded with GL13 K antimicrobial peptide Titanium nanotubes loaded with antimicrobial peptides were placed in a blood dish containing Porphyromonas gingivalis for 48-72 hours under anaerobic conditions.The antimicrobial activity of the materials was evaluated by detecting the concentration of Porphyromonas gingivalis bacterial suspension at different times.Results 1.The surface of smooth titanium sheet was mirror-like,and the titanium nanotubes had a honeycomb-like structure.The diameter of the lumen was fairly uniform,averaging about 80 nm,and the height of the nanotubes was about 2.4 mm.2.The contact angle of the experimental group increased to some extent compared with the blank control group,indicating that the hydrophilicity of the experimental group was weakened;XPS analysis showed that the peak of nitrogen element in the experimental group increased significantly at 400 e V compared with the control group,indicating that GL13 K antimicrobial peptide was successfully fixed on the surface of titanium substrate;FTIR results showed that the peak of the experimental group at 1630 cm-1 was similar to that of the control group.The peak of GL13 K antimicrobial peptide was the same,which indicated that the antimicrobial peptide was successfully fixed on the surface of titanium substrate.The immunofluorescence analysis showed that there was no fluorescence distribution on the surface of the control group.Various fluorescence can be seen on the surface of the experimental group,which confirms that the antimicrobial peptide has successfully immobilized the surface of the titanium dioxide nanotube material.The release curve of antimicrobial peptides showed that the sustained release of antimicrobial peptides was observed on the surface of the self-assembled samples until the 5th day,which indicated that the sustained release effect of the self-assembled group was better than that of the other two groups.3.The antimicrobial coating of titanium dioxide nanotube-GL13 K antimicrobial peptide has certain antimicrobial ability.Conculsion All three methods successfully grafted GL13 K antimicrobial peptide onto the surface of titanium dioxide nanotubes,and successfully constructed antimicrobial coating.Among them,the layer-by-layer self-assembly method has the best sustained release effect,which has a good inhibition effect on Porphyromonas gingivalis and is expected to improve the sustained antimicrobial properties of titanium implant surface. |
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